Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into 11-32A/B,a Widely Used Cytogenic Male Sterile System |
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Authors: | LAI Yun-song, HUANG Hai-qing, XU Meng-yun, WANG Liang-chao, ZHANG Xiao-bo, ZHANG Ji-wen, TU Ju-min |
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Affiliation: | [1]Institute of Crop Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China; [2]Institute of Pomology & Olericulture, Sichuan Agricultural University, Chengdu 611130, P.R. China; [3]Rice Research Institute of Enshi Academy of Agriculture Sciences, Enshi 445000, P.R.China |
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Abstract: | Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis (Bt) δ-endotoxin and provides effective and economic control of lepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR (TAIL-PCR). Bt was located in the promoter region of LOC.Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile (CMS) system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B (maintainer line) and II-32A (male sterile line). Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice. |
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Keywords: | Bt rice male sterile line genome walker marker-assisted backcross (MAB) marker-assisted selection (MAS) SSR |
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